LOCUS       Exported                8787 bp ds-DNA     circular SYN 25-NOV-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pCDHblast-Flx-sORF-Katushka
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8787)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Nov 25, 2021 from SnapGene Viewer 5.0.7
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8787
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    5..122
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        212..415
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     protein_bind    486..519
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     gene            532..1362
                     /label=shuffeled ORF
                     /label=sKatushka
     protein_bind    1375..1408
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     CDS             1432..2139
                     /codon_start=1
                     /product="far-red-shifted derivative of red fluorescent 
                     protein from Entacmaea quadricolor, also known as Katushka 
                     (Shcherbo et al., 2007)"
                     /label=TurboFP635
                     /note="mammalian codon-optimized"
                     /translation="MVGEDSVLITENMHMKLYMEGTVNDHHFKCTSEGEGKPYEGTQTM
                     KIKVVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERITTYEDGG
                     VLTATQDTSLQNGCLIYNVKINGVNFPSNGPVMQKKTLGWEASTEMLYPADSGLRGHSQ
                     MALKLVGGGYLHCSLKTTYRSKKPAKNLKMPGFYFVDRRLERIKEADKETYVEQHEMAV
                     ARYCDLPSKLGHS"
     promoter        2157..2656
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     misc_feature    complement(2192..2215)
                     /label=MSCV R
                     /note="MSCV R"
     misc_feature    2589..2609
                     /label=PGK F
                     /note="PGK F"
     CDS             2678..3076
                     /codon_start=1
                     /label=Blast resistance
                     /note="Blasticidine R"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    3083..3671
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(3554..3565)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3745..3978
                     /label=3' ∂LTR
                     /note="3' ∂LTR"
     polyA_signal    4050..4171
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    complement(4141..4160)
                     /label=EBV-Rev
                     /note="EBV-Rev"
     rep_origin      4190..4325
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     misc_feature    4252..4271
                     /label=SV40PromoterF
                     /note="SV40PromoterF"
     primer_bind     complement(4363..4379)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4387..4403
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(4411..4441)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4456..4477
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4765..5353)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5524..6384)
                     /codon_start=1
                     /label=Amp
                     /note="Ampicilline-R"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6385..6489)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6963..6979
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             6995..7402
                     /label=RSV/5'LTR
                     /note="RSV/5'LTR"
     misc_feature    7446..7571
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7555..7907
                     /label=gag
                     /note="gag"
     misc_feature    8064..8297
                     /label=RRE
                     /note="RRE"
     CDS             8481..8525
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
ORIGIN
        1 taacttttaa aagaaaaggg gggattgggg ggtacagtgc aggggaaaga atagtagaca
       61 taatagcaac agacatacaa actaaagaat tacaaaaaca aattacaaaa attcaaaatt
      121 ttatcgatac tagtattatg cccagtacat gaccttatgg gactttccta cttggcagta
      181 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
      241 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
      301 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
      361 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctcgttt
      421 agtgaaccgt cagatcgcct ggagacgcca tccacgctgt tttgacctcc atagaagatt
      481 ctagaataac ttcgtatagc atacattata cgaagttatg aattcgccac catgaagatt
      541 aacggcgtca acttccctag caacggccct gtgatgcaga agaagaccct gggatgggag
      601 gcaagcacag aaatgctcta cccagctgat agcggactcc gggggcactc ccaaatggct
      661 ctcaagctgg tggggggagg ctacctgcac tgcagcctga agaccacata taggtctaaa
      721 aaaccagcta agaatctgaa gatgcccggc ttctacttcg tcgatagaag gctggagagg
      781 atcaaggagg ccgacaagga aacctatgtg gaacagcacg agatggccgt ggctaggtac
      841 tgcgatctcc cctccaagct cggccattcc atggtgggcg aagacagcgt gctgattacc
      901 gaaaatatgc acatgaaact gtacatggaa ggcaccgtca acgaccacca cttcaaatgc
      961 acctctgagg gggagggcaa gccttatgaa ggcactcaga ccatgaagat caaggtcgtg
     1021 gaagggggac ctctcccatt tgcattcgac atattggcca ctagcttcat gtatggctcc
     1081 aagacattta ttaaccacac acagggcatc cctgatttct tcaaacagtc tttccctgaa
     1141 ggattcacct gggagcggat taccacctac gaggacggag gcgtgctgac agccacccag
     1201 gacacatccc tgcaaaacgg atgtctcatc tataacgtcg gagtgctgac agctacccag
     1261 gatacctccc tgcagaacgg ctgtctgatt tataacgtca agatcaacgg cgtcaacttc
     1321 ccctctaatg ggcctgtcat gcagaagaag acactgggat gataatagct cgagataact
     1381 tcgtatagca tacattatac gaagttatgg atccatctag catgatcaac catggtgggt
     1441 gaggatagcg tgctgatcac cgagaacatg cacatgaaac tgtacatgga gggcaccgtg
     1501 aacgaccacc acttcaagtg cacatccgag ggcgaaggca agccctacga gggcacccag
     1561 accatgaaga tcaaggtggt cgagggcggc cctctcccct tcgccttcga catcctggct
     1621 accagcttca tgtacggcag caaaaccttt atcaaccaca cccagggcat ccccgacttc
     1681 tttaagcagt ccttccctga gggcttcaca tgggagagga tcaccacata cgaagacggg
     1741 ggcgtgctga ccgctaccca ggacaccagc ctccagaacg gctgcctcat ctacaacgtc
     1801 aagatcaacg gggtgaactt cccatccaac ggccctgtga tgcagaagaa aacactcggc
     1861 tgggaggcca gcaccgagat gctgtacccc gctgacagcg gcctgagagg ccatagccag
     1921 atggccctga agctcgtggg cgggggctac ctgcactgct ccctcaagac cacatacaga
     1981 tccaagaaac ccgctaagaa cctcaagatg cccggcttct acttcgtgga caggagactg
     2041 gaaagaatca aggaggccga caaagagacc tacgtcgagc agcacgagat ggctgtggcc
     2101 aggtactgcg acctgcctag caaactgggg cacagctaag cggccgcaat tctaccgggt
     2161 aggggaggcg cttttcccaa ggcagtctgg agcatgcgct ttagcagccc cgctgggcac
     2221 ttggcgctac acaagtggcc tctggcctcg cacacattcc acatccaccg gtaggcgcca
     2281 accggctccg ttctttggtg gccccttcgc gccaccttct actcctcccc tagtcaggaa
     2341 gttccccccc gccccgcagc tcgcgtcgtg caggacgtga caaatggaag tagcacgtct
     2401 cactagtctc gtgcagatgg acagcaccgc tgagcaatgg aagcgggtag gcctttgggg
     2461 cagcggccaa tagcagcttt gctccttcgc tttctgggct cagaggctgg gaaggggtgg
     2521 gtccgggggc gggctcaggg gcgggctcag gggcggggcg ggcgcccgaa ggtcctccgg
     2581 aggcccggca ttctgcacgc ttcaaaagcg cacgtctgcc gcgctgttct cctcttcctc
     2641 atctccgggc ctttcgacct gcagccaatt ggccaccatg gccaagcctt tgtctcaaga
     2701 agaatccacc ctcattgaaa gagcaacggc tacaatcaac agcatcccca tctctgaaga
     2761 ctacagcgtc gccagcgcag ctctctctag cgacggccgc atcttcactg gtgtcaatgt
     2821 atatcatttt actgggggac cttgtgcaga actcgtggtg ctgggcactg ctgctgctgc
     2881 ggcagctggc aacctgactt gtatcgtcgc gatcggaaat gagaacaggg gcatcttgag
     2941 cccctgcgga cggtgccgac aggtgcttct cgatctgcat cctgggatca aagccatagt
     3001 gaaggacagt gatggacagc cgacggcagt tgggattcgt gaattgctgc cctctggtta
     3061 tgtgtgggag ggctaagtcg acaatcaacc tctggattac aaaatttgtg aaagattgac
     3121 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     3181 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     3241 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     3301 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     3361 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     3421 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc
     3481 atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     3541 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     3601 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     3661 cgcctccccg cctggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc
     3721 actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga aaataagatc
     3781 tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
     3841 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
     3901 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
     3961 tgtggaaaat ctctagcagt agtagttcat gtcatcttat tattcagtat ttataacttg
     4021 caaagaaatg aatatcagag agtgagagga acttgtttat tgcagcttat aatggttaca
     4081 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     4141 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gctctagcta tcccgcccct
     4201 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     4261 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     4321 aggcctagac ttttgcagag acggcccaaa ttcgtaatca tggtcatagc tgtttcctgt
     4381 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     4441 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     4501 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     4561 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     4621 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     4681 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     4741 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     4801 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     4861 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     4921 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     4981 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     5041 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     5101 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     5161 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     5221 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     5281 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     5341 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     5401 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     5461 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     5521 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     5581 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     5641 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     5701 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     5761 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     5821 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     5881 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     5941 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     6001 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     6061 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     6121 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     6181 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     6241 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     6301 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     6361 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     6421 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     6481 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
     6541 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga
     6601 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
     6661 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
     6721 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga
     6781 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct
     6841 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
     6901 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
     6961 ttgtaaaacg acggccagtg ccaagctgac gcgtgtagtc ttatgcaata ctcttgtagt
     7021 cttgcaacat ggtaacgatg agttagcaac atgccttaca aggagagaaa aagcaccgtg
     7081 catgccgatt ggtggaagta aggtggtacg atcgtgcctt attaggaagg caacagacgg
     7141 gtctgacatg gattggacga accactgaat tgccgcattg cagagatatt gtatttaagt
     7201 gcctagctcg atacaataaa cgggtctctc tggttagacc agatctgagc ctgggagctc
     7261 tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa
     7321 gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag
     7381 tcagtgtgga aaatctctag cagtggcgcc cgaacaggga cctgaaagcg aaagggaaac
     7441 cagagctctc tcgacgcagg actcggcttg ctgaagcgcg cacggcaaga ggcgaggggc
     7501 ggcgactggt gagtacgcca aaaattttga ctagcggagg ctagaaggag agagatgggt
     7561 gcgagagcgt cagtattaag cgggggagaa ttagatcgcg atgggaaaaa attcggttaa
     7621 ggccaggggg aaagaaaaaa tataaattaa aacatatagt atgggcaagc agggagctag
     7681 aacgattcgc agttaatcct ggcctgttag aaacatcaga aggctgtaga caaatactgg
     7741 gacagctaca accatccctt cagacaggat cagaagaact tagatcatta tataatacag
     7801 tagcaaccct ctattgtgtg catcaaagga tagagataaa agacaccaag gaagctttag
     7861 acaagataga ggaagagcaa aacaaaagta agaccaccgc acagcaagcg gccactgatc
     7921 ttcagacctg gaggaggaga tatgagggac aattggagaa gtgaattata taaatataaa
     7981 gtagtaaaaa ttgaaccatt aggagtagca cccaccaagg caaagagaag agtggtgcag
     8041 agagaaaaaa gagcagtggg aataggagct ttgttccttg ggttcttggg agcagcagga
     8101 agcactatgg gcgcagcctc aatgacgctg acggtacagg ccagacaatt attgtctggt
     8161 atagtgcagc agcagaacaa tttgctgagg gctattgagg cgcaacagca tctgttgcaa
     8221 ctcacagtct ggggcatcaa gcagctccag gcaagaatcc tggctgtgga aagataccta
     8281 aaggatcaac agctcctggg gatttggggt tgctctggaa aactcatttg caccactgct
     8341 gtgccttgga atgctagttg gagtaataaa tctctggaac agattggaat cacacgacct
     8401 ggatggagtg ggacagagaa attaacaatt acacaagctt aatacactcc ttaattgaag
     8461 aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt ggaattagat aaatgggcaa
     8521 gtttgtggaa ttggtttaac ataacaaatt ggctgtggta tataaaatta ttcataatga
     8581 tagtaggagg cttggtaggt ttaagaatag tttttgctgt actttctata gtgaatagag
     8641 ttaggcaggg atattcacca ttatcgtttc agacccacct cccaaccccg aggggacccg
     8701 acaggcccga aggaatagaa gaagaaggtg gagagagaga cagagacaga tccattcgat
     8761 tagtgaacgg atctcgacgg tatcggt
//
